570 Biowissenschaften; Biologie
Refine
Year of publication
- 2017 (212) (remove)
Document Type
- Article (131)
- Doctoral Thesis (64)
- Part of Periodical (6)
- Preprint (3)
- Part of a Book (2)
- Conference Proceeding (2)
- Contribution to a Periodical (2)
- Book (1)
- Working Paper (1)
Is part of the Bibliography
- no (212)
Keywords
- Research article (5)
- Biochemistry (4)
- mitochondria (4)
- Biophysics and structural biology (3)
- Cell biology (3)
- Europe (3)
- LTP (3)
- aging (3)
- hippocampus (3)
- Biodiversität (2)
Institute
- Biowissenschaften (81)
- Biochemie und Chemie (36)
- Institut für Ökologie, Evolution und Diversität (27)
- Exzellenzcluster Makromolekulare Komplexe (25)
- Biochemie, Chemie und Pharmazie (16)
- Senckenbergische Naturforschende Gesellschaft (12)
- Medizin (10)
- Präsidium (10)
- Biodiversität und Klima Forschungszentrum (BiK-F) (9)
- Frankfurt Institute for Advanced Studies (FIAS) (8)
The yeast Rcf1 protein is a member of the conserved family of proteins termed the hypoxia-induced gene (domain) 1 (Hig1 or HIGD1) family. Rcf1 interacts with components of the mitochondrial oxidative phosphorylation system, in particular the cytochrome bc1 (complex III)-cytochrome c oxidase (complex IV) supercomplex (termed III-IV) and the ADP/ATP carrier proteins. Rcf1 plays a role in the assembly and modulation of the activity of complex IV; however, the molecular basis for how Rcf1 influences the activity of complex IV is currently unknown. Hig1 type 2 isoforms, which include the Rcf1 protein, are characterized in part by the presence of a conserved motif, (Q/I)X3(R/H)XRX3Q, termed here the QRRQ motif. We show that mutation of conserved residues within the Rcf1 QRRQ motif alters the interactions between Rcf1 and partner proteins and results in the destabilization of complex IV and alteration of its enzymatic properties. Our findings indicate that Rcf1 does not serve as a stoichiometric component, i.e. as a subunit of complex IV, to support its activity. Rather, we propose that Rcf1 serves to dynamically interact with complex IV during its assembly process and, in doing so, regulates a late maturation step of complex IV. We speculate that the Rcf1/Hig1 proteins play a role in the incorporation and/or remodeling of lipids, in particular cardiolipin, into complex IV and. possibly, other mitochondrial proteins such as ADP/ATP carrier proteins.
Ubiquitination is a widespread post-translational modification that controls multiple steps in autophagy, a major lysosome-mediated intracellular degradation pathway. A variety of ubiquitin chains are attached as selective labels on protein aggregates and dysfunctional organelles, thus promoting their autophagy-dependent degradation. Moreover, ubiquitin modification of autophagy regulatory components is essential to positively or negatively regulate autophagy flux in both non-selective and selective pathways. We review the current findings that elucidate the components, timing, and kinetics of the multivalent role of ubiquitin signals in control of amplitude and selectivity of autophagy pathways as well as their impact on the development of human diseases.
The interaction between the Heat Shock Proteins 70 and 40 is at the core of the ATPase regulation of the chaperone machinery that maintains protein homeostasis. However, the structural details of the interaction remain elusive and contrasting models have been proposed for the transient Hsp70/Hsp40 complexes. Here we combine molecular simulations based on both coarse-grained and atomistic models with coevolutionary sequence analysis to shed light on this problem by focusing on the bacterial DnaK/DnaJ system. The integration of these complementary approaches resulted in a novel structural model that rationalizes previous experimental observations. We identify an evolutionarily conserved interaction surface formed by helix II of the DnaJ J-domain and a structurally contiguous region of DnaK, involving lobe IIA of the nucleotide binding domain, the inter-domain linker, and the β-basket of the substrate binding domain.
Compared to sequence analyses, phylogenetic reconstruction from transposable elements (TEs) offers an additional perspective to study evolutionary processes. However, detecting phylogenetically informative TE insertions requires tedious experimental work, limiting the power of phylogenetic inference. Here, we analyzed the genomes of seven bear species using high throughput sequencing data to detect thousands of TE insertions. The newly developed pipeline for TE detection called TeddyPi (TE detection and discovery for Phylogenetic Inference) obtained 150,513 high-quality TE insertions in the genomes of ursine and tremarctine bears. By integrating different TE insertion callers and using a stringent filtering approach, the TeddyPi pipeline produced highly reliable TE insertion calls, which were confirmed by extensive in vitro validation experiments. Screening for single nucleotide substitutions in the flanking regions of the TEs show that these substitutions correlate with the phylogenetic signal from the TE insertions. Our phylogenomic analyses show that TEs are a major driver of genomic variation in bears and enabled phylogenetic reconstruction of a well-resolved species tree, even with strong signals for incomplete lineage sorting and introgression. The analyses show that the Asiatic black, sun and sloth bear form a monophyletic clade. TeddyPi is open source and can be adapted to various TE and structural variation callers. The pipeline makes it easy to confidently extract thousands of TE insertions even from low coverage genomes of non-model organisms, opening new possibilities for biologists to study phylogenies, evolutionary processes as well as rates and patterns of (retro-)transposition and structural variation.
There is increasing evidence that rapid phenotypic adaptation of quantitative traits is not uncommon in nature. However, the circumstances under which rapid adaptation of polygenic traits occurs are not yet understood. Building on previous concepts of soft selection, i.e. frequency and density dependent selection, I developed and tested the hypothesis that adaptation speed of a polygenic trait depends on the number of offspring per breeding pair in a randomly mating diploid population.
Using individual based modelling on a range of offspring per parent (2–200) in populations of various size (100–10000 individuals), I could show that the by far largest proportion of variance (42%) was explained by the offspring number, regardless of genetic trait architecture (10–50 loci, different locus contribution distributions). In addition, it was possible to identify the majority of the responsible loci and account for even more of the observed phenotypic change with a moderate population size.
The simulation results suggest that offspring numbers may a crucial factor for the adaptation speed of quantitative loci. Moreover, as large offspring numbers translates to a large phenotypic variance in the offspring of each parental pair, this genetic bet hedging strategy increases the chance to contribute to the next generation in unpredictable environments.
Mutations are the ultimate basis of evolution, yet their occurrence rate is known only for few species. We directly estimated the spontaneous mutation rate and the mutational spectrum in the non-biting midge C. riparius with a new approach. Individuals from ten mutation accumulation lines over five generations were deep genome sequenced to count de novo mutations (DNMs) that were not present in a pool of F1 individuals, representing parental genotypes. We identified 51 new single site mutations of which 25 were insertions or deletions and 26 single point mutations. This shift in the mutational spectrum compared to other organisms was explained by the high A/T content of the species. We estimated a haploid mutation rate of 2.1 x 10−9 (95% confidence interval: 1.4 x 10−9 – 3.1 x 10−9) which is in the range of recent estimates for other insects and supports the drift barrier hypothesis. We show that accurate mutation rate estimation from a high number of observed mutations is feasible with moderate effort even for non-model species.
Biodiversität
(2017)
'Biodiversität' ist ein Schlüsselbegriff unserer Zeit, auf dem Forschungsprogramme, ethische Debatten zum Mensch-Natur-Verhältnis und politische Aktivitäten basieren. In der öffentlichen und politischen Kommunikation funktioniert der Begriff offenbar gut. Er transportiert Achtung und Verantwortung für die Natur, Toleranz gegenüber dem Fremden, Freude an der Heterogenität und Mannigfaltigkeit. Biodiversität steht parallel zur kulturellen Vielfalt und passt in unsere durch Pluralismen geprägte Gegenwart. Denn der Begriff drückt nicht nur Enthierarchisierung und Pluralisierung der Perspektiven aus, Verzicht auf eine übergreifende, durchgängig gültige Ordnung und den Eigensinn und Eigenwert jedes einzelnen, auch nichtmenschlichen Wesens. Er steht auch für das Zusammenführen von wissenschaftlichen mit ethischen, ästhetischen und ökonomischen Aspekten eines Gegenstands und für die Hoffnung auf den letztlich harmonischen Zusammenklang des vielstimmigen Mit- und Gegeneinanders.
Glucose homeostasis is tightly regulated by insulin production from ß-cells and glucagon production from α-cells. Changes in the balance of these hormones lead to Diabetes Mellitus (DM), which is foreseen to be the 7th leading cause of death by 2030, warranting a high demand to identify new therapeutics. DM is characterized by a reduction in ß-cell mass and reduced insulin production from ß-cells. α-cell development and fate mainly depend on the activity of the homeodomain-containing transcription factor Aristaless related homeobox (Arx). Conditional loss- of- function of Arx in α-cells leads to their conversion into functional insulin-producing ß-cells and thus an expansion of ß-cell mass. Therefore, inhibition of Arx is an interesting target for the expansion of ß-cells. The zebrafish model provides a fast, cost-effective and reliable translational platform for drug discovery in an in vivo setting. Here, we screened ~6217 small molecules on a transgenic zebrafish line (TgBAC(arxa:Luc2)) in which the arx promoter drives the expression of the luciferase gene which allows a sensitive and quantitative readout of promoter activity. Small molecule screening allowed us to identify 36 candidate repressors of arxa promoter activity. Furthermore, we started to validate these candidates in other assays. Preliminary results showed that DMAT (a potent CK2 inhibitor) and CNS-1102 (NMDA receptor inhibitor) increase functional ß-cell regeneration. By lineage tracing α-cells during ß-cell regeneration, we could show that both DMAT and CNS-1102 promote α- to ß-cell transdifferentiation. Here, we propose that Casein kinase II and NMDA receptor as potential molecular targets that could be exploited for the treatment of diabetes by generating functional beta-cells from the non-beta-cell progenitor, particularly alpha-cells in situ.
Mutualistic interactions between plants and animals can affect both plant and animal communities, and potentially leave imprints on plant demography. Yet, no study has simultaneously tested how trait variation in plant resources shapes the diversity of animal consumers, and how these interactions influence seedling recruitment. Here, we analyzed whether (i) phylogenetic diversity and functional diversity of fruiting plants were correlated with the corresponding diversity of frugivorous birds, and (ii) whether phylogenetic diversity and functional identity of plant and bird communities influenced the corresponding diversity and identity of seedling communities. We recorded mutualistic interactions between fleshy-fruited plants and frugivorous birds and seedling communities in 10 plots along an elevational gradient in the Colombian Andes. We built a phylogeny for plants/seedlings and birds and measured relevant morphological plant and bird traits that influence plant-bird interactions and seedling recruitment. We found that phylogenetic diversity and functional diversity of frugivorous birds were positively associated with the corresponding diversities of fruiting plants, consistent with a bottom-up effect of plants on birds. Moreover, the phylogenetic diversity of seedlings was related to the phylogenetic diversity of plants, but was unrelated to the phylogenetic diversity of frugivorous birds, suggesting that top-down effects of animals on seedlings were weak. Mean seed mass of seedling communities was positively associated with the mean fruit mass of plants, but was not associated with the mean avian body mass in the frugivore communities. Our study shows that variation in the traits of fleshy-fruited plants was associated with the diversity of frugivorous birds and affected the future trajectory of seedling recruitment, whereas the morphological traits of animal seed dispersers were unrelated to the phylogenetic and functional structure of seedling communities. These findings suggest that bottom-up effects are more important than top-down effects for seed-dispersal interactions and seedling recruitment in diverse tropical communities.
Morphological malformations induced by tributyltin (TBT) exposure during embryonic development have already been characterized in various taxonomic groups, but, nonetheless, the molecular processes underlying these changes remain obscure. The present study provides the first genome-wide screening for differentially expressed genes that are linked to morphological alterations of gonadal tissue from chicken embryos after exposure to TBT. We applied a single injection of TBT (between 0.5 and 30 pg as Sn/g egg) into incubated fertile eggs to simulate maternal transfer of the endocrine disruptive compound. Methyltestosterone (MT) served as a positive control (30 pg/g egg). After 19 days of incubation, structural features of the gonads as well as genome-wide gene expression profiles were assessed simultaneously. TBT induced significant morphological and histological malformations of gonadal tissue from female embryos that show a virilization of the ovaries. This phenotypical virilization was mirrored by altered expression profiles of sex-dependent genes. Among these are several transcription and growth factors (e.g. FGF12, CTCF, NFIB), whose altered expression might serve as a set of markers for early identification of endocrine active chemicals that affect embryonic development by transcriptome profiling without the need of elaborate histological analyses.
Die vorliegende Arbeit befasst sich mit der Entwicklung und Validierung eines Messinstrumentes zur Erfassung von psychopathischen Persönlichkeitseigenschaften, dem Fragebogen Psychopathischer Persönlichkeitseigenschaften (FPP), mit der Entwicklung eines Messinstrumentes, das antisoziales Verhalten und Kriminalität getrennt erfasst, der Checkliste für antisoziale Verhaltensweisen und Kriminalität (CAV/K) und mit der differenzierten Untersuchung der Vorhersage von Kriminalität durch psychopathische Eigenschaften.
Psychopathie ist eines der wichtigsten Konstrukte in der Vorhersage von kriminellem und antisozialem Verhalten (Hemphill, Hare & Wong, 1998). Die Konzeption der psychopathischen Persönlichkeit stellt affektive und interpersonelle Eigenschaften in den Vordergrund und definiert Kriminalität als Konsequenz statt Kernsymptom der Psychopathie (Skeem & Cooke, 2010a). Bisherige Ergebnisse zur prädiktiven Validität von psychopathischen Eigenschaften hinsichtlich krimineller Rückfälligkeit weisen allerdings darauf hin, dass affektive und interpersonelle Facetten der Psychopathie keinen Erklärungswert über eine kriminelle Vorgeschichte hinaus haben (Eher, Schilling, Mönichweger, Haubner-MacLean & Rettenberger, 2012; Walters, Knight, Grann & Dahle, 2008; Walters, Wilson & Glover, 2011). Des Weiteren wird im Rahmen der adaptiven Psychopathie postuliert, dass psychopathische Eigenschaften auch hoch ausgeprägt sein können, ohne dass sich die Personen kriminell verhalten (Hall & Benning, 2006). Ergebnisse aus einer Metananalyse eines etablierten Fragebogens zur Psychopathie, dem Psychopathic Personality Inventory (PPI; Lilienfeld & Andrews, 1996) ergaben, dass der erste von zwei Faktoren nicht mit Indizes antisozialer Verhaltensweisen korrelieren. Diese Ergebnisse sprechen dafür, dass der Zusammenhang zwischen Psychopathie und Kriminalität genauer untersucht werden muss, da ein Fehlen dieser Assoziation das Konstrukt Psychopathie obsolet erscheinen lässt.
Voraussetzung für die valide Untersuchung dieses Zusammenhangs ist die Verfügbarkeit von psychometrisch hochwertigen und validen Instrumenten zur Erfassung der Variablen. Die Erfassung von psychopathischen Eigenschaften im Selbstbericht ist ökonomisch und Beurteilungen fallen valide aus (Ray et al., 2013). Der Fragebogen PPI (Lilienfeld & Andrews, 1996) ist bereits gut etabliert, weist jedoch einige methodische Schwächen auf, wie z. B. eine Vielzahl von Items sowie eine schwer replizierbare Faktorenstruktur.
Deshalb wurde im ersten Schritt der Fragebogen Psychopathischer Persönlichkeitseigenschaften (FPP, Etzler & Rohrmann, 2017a, 2017b) entwickelt, der die Kritikpunkte des PPI überwindet und psychopathische Persönlichkeitseigenschaften mit hoher psychometrischer Gute misst. Der FPP erfasst sechs Facetten der Psychopathie, die theoretisch hergeleitet wurden: Fehlende Empathie, Furchtlosigkeit, Narzisstischer Egozentrismus, Impulsivität, Soziale Manipulation und Macht. Diese Facetten unterscheiden sich bis auf Furchtlosigkeit von den Skalen des PPI. Die Items sind kurz und inhaftierungsadäquat formuliert und weisen eine mittlere Situationsspezifität auf. Die Itemselektion basierte auf einer Stichprobe von n = 173 Straftätern und n = 132 Zivilpersonen, was repräsentativ für den Anwendungsbereich des FPP ist. Ergebnisse weisen auf gute psychometrische Eigenschaften, wie Itemkennwerte und Reliabilität (ω = .85) hin, auf überwiegend gleiche Messeigenschaften in Zivil- und Haftstichprobe sowie auf eine Konvergenz mit dem PPI (r = .677∗∗) in erwarteter Höhe. Erwartungskonforme Zusammenhänge mit weiteren Konstrukten, Korrelationen mit antisozialem Verhalten, Disziplinarmaßnahmen in Haft und Inhaftierung selbst unterstützen die Kriteriumsvalidität des FPP.
Die Vorhersage von Kriminalität setzt des Weiteren eine präzise Definition ebendieser Variable voraus. Allerdings wurde Kriminalität bis dato in den publizierten Studien uneinheitlich definiert und operationalisiert. Im Rahmen der vorliegenden Arbeit wurde Kriminalität als soziologisches Konstrukt definiert, das maßgeblich von der Gesetzgebung des Landes und weiteren Faktoren abhängt, die außerhalb einer Person liegen (Eysenck, 1977). Demgegenüber wurde antisoziales Verhalten psychologisch definiert, als Verhalten, das anderen Personen schadet aber dem Akteur selbst von Vorteil ist. Da für die Erfassung von antisozialem Verhalten und Kriminalität bis dato noch kein Messinstrument vorliegt, wurde die Checkliste für Antisoziale Verhaltensweisen und Kriminalität (CAV/K, Etzler & Rohrmann, 2017a) entwickelt, die beide Variablen getrennt voneinander aber dennoch simultan erfasst.
Nach Bereitstellung der Definitionen und Messinstrumente wurde untersucht, wie Kriminalität durch psychopathische Eigenschaften vorhergesagt werden kann (Etzler, Rettenberger & Rohrmann, eingereicht). Im Rahmen der adaptiven Psychopathie wurde angenommen, dass Moderatorvariablen den Zusammenhang zwischen Psychopathie und Kriminalität beeinflussen, wobei verbale Intelligenz hierbei eine zentrale Rolle spielt (Wall, Sellbom & Goodwin, 2013). Antisoziales Verhalten und Kriminalität (als offizielle Verurteilung) wurden getrennt voneinander definiert. Es zeigte sich im Rahmen eines moderierten Mediationsmodells, dass psychopathische Eigenschaften antisoziales Verhalten mit hoher Präzision vorhersagen konnten, insbesondere war die Wahrscheinlichkeit, dafür verurteilt zu werden und damit offiziell kriminell zu sein höher für Personen mit niedriger verbaler Intelligenz als für Personen mit höherer Intelligenz, da für verbale Intelligenz ein protektiver Effekt hinsichtlich Kriminalität gefunden wurde.
Die vorliegende Studie verfolgt einen integrativen Ansatz, indem Eigenschaften untersucht werden, die bei Straftätern und Zivilpersonen gleichermaßen vorliegen. Der FPP kann in der Forschung als ökonomisches Instrument eingesetzt werden sowie als Screeninginstrument in der Praxis, z. B. für die Planung des Vollzugsverlaufs in Haft. Anhand der Ergebnisse des FPP können antisoziale Verhaltensweisen in vielen Kontexten (z. B. Mobbing in Unternehmen, deviantes Verhalten in Haft) vorhergesagt werden und unter Hinzunahme weiterer Moderatoren, wie etwa verbale Intelligenz, kann eine präzisere Vorhersage von Kriminalität anhand psychopathischer Eigenschaften erfolgen.
Neurleptic drugs, e.g., aripiprazole, targeting the dopamine D2S and D3 receptors (D2SR and D3R) in the central nervous system are widely used in the treatment of several psychotic and neurodegenerative diseases. Therefore, a new series of benzothiazole-based ligands (3-20) was synthesized by applying the bioisosteric approach derived from the selective D3Rs ligand BP-897 (1) and its structurally related benz[d]imidazole derivative (2). Herein, introduction of the benzothiazole moiety was well tolerated by D2SR and D3R binding sites leading to antagonist affinities in the low nanomolar concentration range at both receptor subtypes. However, all novel compounds showed lower antagonist affinity to D3R when compared to that of 1. Further exploration of different substitution patterns at the benzothiazole heterocycle and the basic 4-phenylpiperazine resulted in the discovery of high dually acting D2SR and D3R ligands. Moreover, the methoxy substitution at 2-position of 4-phenylpiperazine resulted in significantly (22-fold) increased D2SR binding affinity as compared to the parent ligand 1, and improved physicochemical and drug-likeness properties of ligands 3-11. However, the latter structural modifications failed to improve the drug-able properties in ligands having un-substituted 4-phenylpiperazine analogs (12-20). Accordingly, compound 9 showed in addition to high dual affinity at the D2SR and D3R [Ki (hD2SR) = 2.8 ± 0.8 nM; Ki (hD3R) = 3.0 ± 1.6 nM], promising clogS, clogP, LE (hD2SR, hD3R), LipE (hD2SR, hD3R), and drug-likeness score values of −4.7, 4.2, (0.4, 0.4), (4.4, 4.3), and 0.7, respectively. Also, the deaminated analog 10 [Ki (hD2SR) = 3.2 ± 0.4 nM; Ki (hD3R) = 8.5 ± 2.2 nM] revealed clogS, clogP, LE (hD2SR, hD3R), LipE (hD2SR, hD3R) and drug-likeness score values of −4.7, 4.2, (0.4, 0.4), (3.9, 3.5), and 0.4, respectively. The results observed for the newly developed benzothiazole-based ligands 3-20 provide clues for the diversity in structure activity relationships (SARs) at the D2SR and D3R subtypes.
The synergetic effects of combining structural biology and epr spectroscopy on membrane proteins
(2017)
Protein structures as provided by structural biology such as X-ray crystallography, cryo-electron microscopy and NMR spectroscopy are key elements to understand the function of a protein on the molecular level. Nonetheless, they might be error-prone due to crystallization artifacts or, in particular in case of membrane-imbedded proteins, a mostly artificial environment. In this review, we will introduce different EPR spectroscopy methods as powerful tools to complement and validate structural data gaining insights in the dynamics of proteins and protein complexes such that functional cycles can be derived. We will highlight the use of EPR spectroscopy on membrane-embedded proteins and protein complexes ranging from receptors to secondary active transporters as structural information is still limited in this field and the lipid environment is a particular challenge.
Telomeric G-quadruplexes have recently emerged as drug targets in cancer research. Herein, we present the first NMR structure of a telomeric DNA G-quadruplex that adopts the biologically relevant hybrid-2 conformation in a ligand-bound state. We solved the complex with a metalorganic gold(III) ligand that stabilizes G-quadruplexes. Analysis of the free and bound structures reveals structural changes in the capping region of the G-quadruplex. The ligand is sandwiched between one terminal G-tetrad and a flanking nucleotide. This complex structure involves a major structural rearrangement compared to the free G-quadruplex structure as observed for other G-quadruplexes in different conformations, invalidating simple docking approaches to ligand-G-quadruplex structure determination
The impact of the incorporation of a non-natural amino acid (NNAA) on protein structure, dynamics, and ligand binding has not been studied rigorously so far. NNAAs are regularly used to modify proteins post-translationally in vivo and in vitro through click chemistry. Herein, structural characterisation of the impact of the incorporation of azidohomoalanine (AZH) into the model protein domain PDZ3 is examined by means of NMR spectroscopy and X-ray crystallography. The structure and dynamics of the apo state of AZH-modified PDZ3 remain mostly unperturbed. Furthermore, the binding of two PDZ3 binding peptides are unchanged upon incorporation of AZH. The interface of the AZH-modified PDZ3 and an azulene-linked peptide for vibrational energy transfer studies has been mapped by means of chemical shift perturbations and NOEs between the unlabelled azulene-linked peptide and the isotopically labelled protein. Co-crystallisation and soaking failed for the peptide-bound holo complex. NMR spectroscopy, however, allowed determination of the protein-ligand interface. Although the incorporation of AZH was minimally invasive for PDZ3, structural analysis of NNAA-modified proteins through the methodology presented herein should be performed to ensure structural integrity of the studied target.
Feeding type and development drive the ingestion of microplastics by freshwater invertebrates
(2017)
Microscopic plastic items (microplastics) are ubiquitously present in aquatic ecosystems. With decreasing size their availability and potential to accumulate throughout food webs increase. However, little is known on the uptake of microplastics by freshwater invertebrates. To address this, we exposed species with different feeding strategies to 1, 10 and 90 µm fluorescent polystyrene spheres (3–3 000 particles mL−1). Additionally, we investigated how developmental stages and a co-exposure to natural particles (e.g., food) modulate microplastic ingestion. All species ingested microplastics in a concentration-dependent manner with Daphnia magna consuming up to 6 180 particles h−1, followed by Chironomus riparius (226 particles h−1), Physella acuta (118 particles h−1), Gammarus pulex (10 particles h−1) and Lumbriculus variegatus (8 particles h−1). D. magna did not ingest 90 µm microplastics whereas the other species preferred larger microplastics over 1 µm in size. In C. riparius and D. magna, size preference depended on the life stage with larger specimens ingesting more and larger microplastics. The presence of natural particles generally reduced the microplastics uptake. Our results demonstrate that freshwater invertebrates have the capacity to ingest microplastics. However, the quantity of uptake depends on their feeding type and morphology as well as on the availability of microplastics.
Infections with multidrug resistant bacterial strains like Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa or Acinetobacter baumanii that can accumulate resistance mechanisms against different groups of drugs cause increasing problems for the health care system. Multidrug efflux pumps are able to transport different classes of substances, providing a basic resistance to different antibiotics. Especially when they are overexpressed they can keep bacterial cells alive under antibiotic pressure unless other high level resistance mechanisms like expression of β-lactamases are established. One example for a clinically relevant multidrug efflux pump is the AcrAB/TolC tripartite system of E. coli, that transports a variety of different substrates, including besides antibiotics dyes, detergents, bile salts and organic compounds from the periplasm or the inner membrane out of the cell. AcrB is the inner membrane component of the protein complex that determines not only the substrate specificity of the tripartite system but energises the transport through the whole system process via proton transduction as well. TolC is the outer membrane spanning protein that forms a pore in the outer membrane enabling the system to transport drugs over the latter out of the cell. The periplasmic membrane fusion protein AcrA connects AcrB and TolC in the periplasm completing the channel from the periplasm, respective the inner membrane to the extracellular space. AcrB assembles as trimers, in asymmetric crystal structures each of the protomers adapts a different conformation designated L(oose), T(ight) and O(pen). In the protomers tunnels open up and collaps in different conformations. In the L protomer a periplasmic cleft opens up that can initially bind substrates to the periplasmic part of AcrB. In the T conformation the deep binding pocket opens that is assumed to bind substrates tightly that were bound to the access pocket before. As well in the T conformation a second pathway leading to the deep binding pocket opens that can guide substrates from a groove between transmembrane helices TM7, TM8 and TM9, the TM8 groove, that is connected with socalled tunnel 1 that ends in the deep binding pocket. In the O conformation a new tunnel opens that connects the collapsing deep binding pocket with the periplasmic space, respective the channel through the periplasmic space formed from AcrA and TolC. Substrates were cocrystallised in access and deep binding pocket verifying their role in substrate transport. In the TM8 groove in high resolution crystal structures DDM molecules were cocrystallised in L and T conformation, indicating that the AcrB substrate DDM may utilise this entrance to the deep binding pocket. The asymmetry observed in the AcrB trimers trongly suggests a peristaltic pump mechanism. The functional rotation cycle demands communication between the subunits and tight control of substrate load of protomers during the transport to optimise the ration between protons that are transduced and substrates transported. Indeed it was shown that AcrB transport mechanism is positively cooperative for some β-lactam substrates. For the communication between the subunits it was assumed that ionic interaction between ion pairs established between charged amino acids at the interfaces of protomers in different conformations are of special importance. Thus the amino acids engaged in ionic interactions, respective ion pairs D73-K131, E130-K110, D174-K110, R168, R259-E734 were substituted with non-charged amino acids pairwise and phenotypes were determined in plate dilution assays and MIC experiments. No evidence for a general, substrate independent, reduction of AcrB activity, that would be expected when the ionic residues are of special importance for AcrB function, could be found with the methods applied. Substitutions were not only combined pairwise according to the putative ion pairs but as well in combinations of R168A with D174N, E130Q and K131M. AcrB activity is reduced for the variant R168A_D174N significantly, activity decreases further for quadruple variant E130Q_K131M_ R168A_D174N. Because the reduced activity is only observed in this combination of substitutions the phenotype must result from accumulation of small effects of the single substitutions. R168A may destabilise the protomer interfaces, as its side chain is oriented in direction to the neighbouring protomer at all interfaces, enhancing substratespecific effects of substitutions E130Q, K131M, D174N that are not in all conformations oriented towards the neighbouring protomer but as well along the substrate transport pathway. Further investigations to figure out the details of the effects observed were not conducted because fluctuating expression of the variants hindered experimental procedures.
In another approach TM8 was in focus of the interest. As mentioned above it is a possible substrate entrance in the inner membrane. The linker between TM8 and the periplasmic PC2 subdomain undergoes a coil-to-helix transition when AcrB cycles through L, T and O conformations. Linking the transmembrane part of AcrB that provides the energy for the transport process via proton transduction with the periplasmic part harbouring the major part of the substrate pathway assignes TM8 and the periplasmic linker (859-876) an important role in the function of AcrB. Thus it was investigated with an alanine-scan of residues 859 to 884 and G/P respective P/G exchange followed by phenotype characterisation in growth curve and plate dilution assays of selected variants. In the phenotype determinations none of the variants, except G861P that seems to cause massive sterical restriction in an α-helical region, displayed a general, substrate independent decrease of AcrB activity. Thus it is concluded that the individual properties of amino acids in TM8 and the periplasmic linker are not of general importance for the mechanism of AcrB. The substitution of individual amino acids had impact on uptake of different substrates in plate dilution assays in a substrate dependent manner. The uptake of some substrates, like erythromycin or chloramphenicol is more affected than that of others with rhodamine 6G resistance being only reduced for the G861P variant. A relation between the PSA of substrates and reduced activity of AcrB was observed. in Substrates with higher PSA values are more affected by substitutions in TM8 or periplasmic linker, resulting in the conclusion that substrates with higher PSA are more likely to be taken up via the TM8 groove/tunnel 1 pathway than those with lower PSA values.
Phytotoxic dioxolanones from Guignardia bidwellii can be described as potential virulence factors which cause the formation of lesions upon an infection by G. bidwellii. The toxin guignardic acid was found in planta of G. bidwellii-infected Vitis vinifera leaves, whereas no phytotoxic dioxolanones were detected in uninfected leaf material. Secondary metabolism analyses of further phytopathogenic fungi from the genus Guignardia led to the observation that all species investigated can produce the phytotoxins known from G. bidwellii. In addition to these studies, it was demonstrated that phenguignardic acid is biosynthetically derived from two molecules of phenylalanine and that phenylalanine is a key precursor in the biosynthesis of the two other phytotoxins – alaguignardic acid and guignardic acid.
Extraembryonic endoderm stem (XEN) cell lines can be derived and maintained in vitro and reflect the primitive endoderm lineage. Platelet-derived growth factor receptor alpha (PDGFRA) is thought to be essential for the derivation and maintenance of mouse XEN cell lines. Here, we have re-evaluated this requirement for PDGFRA. We derived multiple PDGFRA-deficient XEN cell lines from postimplantation and preimplantation embryos of a PDGFRA-GFP knockout strain. We also converted PDGFRA-deficient embryonic stem cell lines into XEN cell lines chemically by transient culturing with retinoic acid and Activin A. We confirmed the XEN profile of our 12 PDGFRA-deficient cell lines by immunofluorescence with various markers, by NanoString gene expression analyses, and by their contribution to the extraembryonic endoderm of chimeric embryos produced by injecting these cells into blastocysts. Thus, PDGFRA is not essential for the derivation and maintenance of XEN cell lines.
Mitochondria are the "power plants" of eukaryotic cells involved cellular energy metabolism and lead the generation of most of the cellular "energy currency" adenosine triphosphate (ATP). In addition, they have other crucial functions including the control of programmed cell death, iron/sulfur cluster biogenesis and copper and calcium homeostasis. Mitochondrial dysfunction is deleterious and leads to degeneration, disease and aging. A number of individual pathways are active in keeping mitochondria functional over longer periods of time and thereby have a strong impact on lifespan. These mitochondrial quality control (mtQC) pathways occur at different molecular and cellular levels and are all limited in their capacity. They do not all work at the same time. Some of them are induced when others fail. Currently, the underlying molecular interaction of pathways and their regulation is only initially elucidated. ...
The objectives of this thesis were to understand how distinct classes of cell types interact to shape oscillatory activity in cortical circuits of the turtle. We chose the turtle cortex as a model system for cortical computations for two reasons. One is that the phylogenetic position of turtles makes their cortex functionally and anatomically particularly interesting. The second is that reptilian brains present several unique experimental advantages. Turtles have a three-layered cortex that forms the dorsalmost part of their pallium and receives direct input from visual thalamus. Thus turtle cortex, while sharing several features with mammalian cortices, constitutes a simpler system for studying cortical computations and dynamics. Freshwater turtles are semiaquatic species, that dive for hours and hibernate for months without breathing. Their brains are adapted to these behaviors so that they can operate under severe anoxia. This property allows for ex vivo wholebrain and whole-cortex (”cortical slab”) preparations in vitro, enabling the use of many sophisticated techniques for monitoring activity in parallel.
I thus set out to utilize the advantages of our model system, by using optogenetic methods to reliably evoke oscillations in an ex vivo whole-cortex preparation while observing activity in parallel with planar multi-electrode arrays (MEA), linear silicon depth-electrodes and patch-clamp recording techniques. This required several technical aspects to be solved. Prior work in turtle cortex (Prechtl, 1994; Prechtl et al., 1997; Senseman and Robbins, 2002) indicated that visual stimuli evoke complex activity patterns (e. g. wave patterns) in dorsal cortex. The goal was to examine these dynamics in detail and to provide mechanistic explanations for them whenever possible. The recent advent of optogenetics, the development of microelectrode arrays, and the possibility to combine these techniques with classical electrophysiological approaches on a resistant, accessible and stable preparation led me to explore a number of technical avenues.
First I had to establish gene delivery methods in reptiles. I settled on recombinant viruses, and show results from several serotypes of adeno-associated virus (AAV), i lentivirus and rabies virus. I report successful gene expression of genes of interest with several subtypes of AAV, including the commonly used AAV2/1 and AAV2/5 serotypes. Second I had to find promoters enabling global and cell-type specific gene expression in reptiles. Ubiquitous high-yield promoters such as CAG/CB7 or CMV drive high levels of expression in turtles; cell-type specific promoters such as hSyn (expression limited to neurons) and CaMKIIa (expression limited exclusively o mostly to excitatory neurons) appear similarly biased in turtles. Other cell-type specific promoters reported in the literature (fNPY, fPV, fSST) failed to express in turtles.
A second major aspect of my work focused on electrophysiological recordings using microelectrode arrays and the interpretation of extracellular signals recorded from cortex in ex vivo preparations. We observed that spike signals produced by pyramidal and inhibitory neurons were very often followed by a slower potential. We identified these slower potentials as reflections of synaptic currents, and thus of the axonal projections of the neurons, at least within the deep layers of cortex. This also resulted in a means to classify neurons as excitatory or inhibitory with much higher reliability than classical methods (e. g. spike width). The final aspect of my work concerns the use of optogenetics to dissect the mechanisms of cortical oscillations and wave propagation. I show that oscillations can be induced by light in turtle cortex after transfection with AAV2/1 carrying the gene for channelrhodopsin 2 (ChR2). By using the CaMKIIa promoter, ChR2 induced currents are limited to LII/III excitatory cells; we can therefore control excitatory drive to cortical networks. If this drive is strong enough, layer III inhibitory interneurons are recruited and fire in a concerted fashion, silencing the excitatory population. The visually evoked 20 Hz oscillations observed in chronically recorded animals (Schneider, 2015) or in anaesthetized animals (Fournier et al., in press) thus appear to result from a feedback loop between E and I cells within layers II & III. Details of these interactions are being investigated but - layer I interneurons, by contrast, do not seem to be involved. By pulsing light I could control the frequency of the oscillations within a range of several Hz around the natural oscillation frequency. Above this range, cortex could only follow the stimulus at a fraction (1/2, 1/3,...) of the light pulse frequency. Using a digital micromirror device, I limited activation of the cortical networks spatially, enabling the study of wave propagation in this system.
Reptilian cortex offers a relatively simple model system for a reductionist and comparative strategy on understanding cortical computations and dynamics. Turtle dorsal cortex could thus give fundamental insights to the primordial organization tional, computational and functional principles of cortical networks. These insights are relevant to our understanding of mammalian brains and may prove valuable to decipher fundamental questions of modern neuroscience.
Colorectal cancer (CRC) has the third highest incidence and the fourth highest mortality rate worldwide and represents a substantial health care burden and affects the life of millions of people. CRC is a genetic disease caused by the stepwise accumulation of genetic alterations. The initiating event in colorectal carcinogenesis is the aberrant activation of the WNT pathway, but other pathways are also commonly deregulated, including the PI3K/AKT pathway. A number of previous studies using genetically engineered mouse models aimed at dissecting the exact role of PI3K/AKT pathway in CRC, but have yielded in rather conflicting results. Despite the inconsistent results, these studies already put forward the idea that PI3K/AKT signaling in combination with other genetic events might substantially contribute to tumor progression. Since the PI3K/AKT pathway is frequently activated in CRC, it represents an ideal candidate for therapeutic intervention. Although extensive efforts had led to the development of numerous inhibitors targeting the PI3K/AKT pathway, the diversity of genetic alterations can challenge the identification of the most effective therapeutic targets. Therefore, the discovery of shared tumor-promoting mechanisms downstream of these genetic alterations might unravel new biomarkers and druggable targets. The aim of this study was to elucidate the precise role of PI3K/AKT pathway during the course of colorectal carcinogenesis and to decipher novel protumorigenic molecular mechanisms downstream of PI3K/AKT activation that can be used for therapeutic intervention.
To obtain a better insight into the role of the PI3K/AKT pathway during colorectal carcinogenesis, mice expressing an oncogenic variant of AKT1 (AktE17K) specifically in the intestinal epithelial cells (IEC) were used. At the age of 6 months untreated AktE17K mice showed clearly perturbed intestinal homeostasis, but no tumor formation. To induce colonic tumorigenesis, AktE17K mice were subjected to treatment with the colonic carcinogen azoxymethane (AOM). In response to AOM, AktE17K mice developed invasive but non-metastatic tumors, which showed strong nuclear accumulation of TP53. To investigate the role of PI3K/AKT signaling specifically in CRC progression, AktE17K mice were crossed to TP53-deficient mice (Tp53ΔIEC). Unlike AktE17K mice, untreated Tp53ΔIEC; AktE17K, developed highly invasive small
intestinal tumors by the age of 6 months. To investigate the role of AKT hyperactivation in colonic tumor progression, Tp53ΔIEC; AktE17K mice were subjected to AOM treatment. AKT hyperactivation significantly enhanced tumor progression and induced metastatic dissemination.
To get a better insight how AKT signaling can promote tumor progression, whole tumor tissues from AOM-treated Tp53ΔIEC and Tp53ΔIEC; AktE17K mice were subjected to next generation mRNA sequencing and phospho-proteomic analysis by mass spectrometry. Both analyses indicated that AKT hyperactivation expands the inflammatory tumor microenvironment and upregulates pathways associated with invasion and metastasis. Importantly, Gene Set Enrichment Analysis revealed that AOM-induced colon tumors of Tp53ΔIEC; AktE17K animals, are highly similar in their gene expression profile to the CMS4 subtype of human CRC, which is associated with worse overall- and relapse-free survival. Gene expression analysis also suggested elevated NOTCH signaling in the Tp53ΔIEC; AktE17K tumors. Interestingly, while the expression of Notch3 mRNA was increased in the tumors of Tp53ΔIEC; AktE17K mice, the expression of the other NOTCH receptors was unaffected by AKT hyperactivation. In vitro experiments using TP53-deficient mouse tumor organoids with hyperactive AKT signaling confirmed the direct, tumor cell-intrinsic link between AKT activation and increased Notch3 expression. Moreover, inhibition of EZH2 mimicked the effect of AKT hyperactivation on Notch3 expression, suggesting that AKT regulates Notch3 via an epigenetic mechanism.
Knock-down of Notch3 in TP53-deficient mouse tumor organoids with hyperactive AKT signaling resulted in differential regulation of several pathways with potential role in invasion and metastasis and in cell death and survival. Subsequent in vivo experiments confirmed the role of NOTCH3 signaling in CRC progression. Treatment of AOM-induced Tp53ΔIEC; AktE17K mice with a NOTCH3 antagonistic antibody or the γ-secretase inhibitor DAPT significantly reduced invasion and metastasis. Importantly, NOTCH3 expression was also found to be associated with human CRC progression, suggesting that NOTCH3 represent a valid target for the treatment of CRC. This work, using genetically engineered mouse models and advanced in vitro techniques, has demonstrated a strong tumor promoting role for PI3K/AKT signaling in CRC progression and has identified NOTCH3 signaling as a potential therapeutic target downstream of the PI3K/AKT pathway.
Fatty acids (FAs) are considered strategically important platform compounds that can be accessed by sustainable microbial approaches. Here we report the reprogramming of chain-length control of Saccharomyces cerevisiae fatty acid synthase (FAS). Aiming for short-chain FAs (SCFAs) producing baker’s yeast, we perform a highly rational and minimally invasive protein engineering approach that leaves the molecular mechanisms of FASs unchanged. Finally, we identify five mutations that can turn baker’s yeast into a SCFA producing system. Without any further pathway engineering, we achieve yields in extracellular concentrations of SCFAs, mainly hexanoic acid (C6-FA) and octanoic acid (C8-FA), of 464 mg l−1 in total. Furthermore, we succeed in the specific production of C6- or C8-FA in extracellular concentrations of 72 and 245 mg l−1, respectively. The presented technology is applicable far beyond baker’s yeast, and can be plugged into essentially all currently available FA overproducing microorganisms.
Three-dimensional multicellular aggregates such as spheroids provide reliable in vitro substitutes for tissues. Quantitative characterization of spheroids at the cellular level is fundamental. We present the first pipeline that provides three-dimensional, high-quality images of intact spheroids at cellular resolution and a comprehensive image analysis that completes traditional image segmentation by algorithms from other fields. The pipeline combines light sheet-based fluorescence microscopy of optically cleared spheroids with automated nuclei segmentation (F score: 0.88) and concepts from graph analysis and computational topology. Incorporating cell graphs and alpha shapes provided more than 30 features of individual nuclei, the cellular neighborhood and the spheroid morphology. The application of our pipeline to a set of breast carcinoma spheroids revealed two concentric layers of different cell density for more than 30,000 cells. The thickness of the outer cell layer depends on a spheroid’s size and varies between 50% and 75% of its radius. In differently-sized spheroids, we detected patches of different cell densities ranging from 5 × 105 to 1 × 106 cells/mm3. Since cell density affects cell behavior in tissues, structural heterogeneities need to be incorporated into existing models. Our image analysis pipeline provides a multiscale approach to obtain the relevant data for a system-level understanding of tissue architecture.
Rhabdomyosarcoma is the most common paediatric soft-tissue sarcoma, and for tumour recurrence, the prognosis is still unfavourable. The current standard therapy consisting of surgery, radiation and combined chemotherapy does not consider the specific biology of this tumour.
Histone deacetylases (HDACs) and the Lysine-specific demethylase-1 (LSD1) are two epigenetic modifiers which are both part of repressor complexes leading to transcriptional silencing of target genes. Whereas HDACs lead to deacetylation of several lysine-residues within the histone tail, LSD1 is specific for demethylation of H3K4me2 and H3K4me1, as well as in a different context for H3K9me2. Rhabdomyosarcoma is reported to harbour high levels of LSD1, but the functional relevance is yet unclear. HDAC inhibition proved to be effective as single agent treatment, however, the proximity of HDAC1/2 and LSD1 in repressor complexes at the DNA implies a suitable rationale for a combination therapy potentially leading to cooperative effects on target gene transcription. In this study, we aimed to evaluate the potential of a combined LSD1 and HDAC inhibition for cell death induction in rhabdomyosarcoma cell lines. Whereas LSD1 inhibitors failed to induce cell death on their own, the combined inhibition of HDACs and LSD1 resulted in highly synergistic cell death induction. This effect extended to several combinations of LSD1 and HDAC inhibitors as well as to four different rhabdomyosarcoma cell lines, two of embryonal and two of alveolar histology.
With the use of the HDAC inhibitor JNJ-26481585 and the reversible LSD1 inhibitor GSK690, we demonstrated that the cell death induced by the combination matches with the details of intrinsic mitochondrial apoptosis. JNJ-26481585/GSK690-induced cell death is partially caspase-dependent and leads to caspase cleavage, followed by substrate cleavage as shown for PARP, as well as loss of the mitochondrial membrane potential.
Furthermore, JNJ-26481585 and GSK690 acted together to transcriptionally upregulate the proapoptotic proteins NOXA, BIM and BMF, which resulted in respective changes on protein level for both cell lines. However, the antiapoptotic BCL-2 family proteins BCL-2, MCL-1 and BCL-xL displayed only minor changes in protein levels upon treatment with GSK690 and JNJ-26481585, which did not rely on transcriptional activity. Therefore, the increase in proapoptotic proteins induces a shift towards proapoptotic signalling at the mitochondrial membrane. This shift is functionally relevant since knockdown of a proapoptotic protein or overexpression of one of the antiapoptotic proteins BCL-2 and MCL-1, as well as a stabilized mutant MCL-1, can significantly protect from GSK690/JNJ-26481585-induced cell death.
Knockdown of the mitochondrial membrane protein BAK, which is directly guarding the mitochondrial membrane integrity, potently protected from GSK690/JNJ-26481585- induced cell death, directly linking the shift in the BCL-2 family proteins to the observed loss of mitochondrial membrane potential and the further downstream activation of caspases. Furthermore, treatment with JNJ-26481585 and GSK690 resulted in a cell cycle arrest in G2/M phase, indicating additional effects on the tumour cells beside apoptosis induction. Taken together, the combined inhibition of LSD1 and HDACs is a promising strategy for rhabdomyosarcoma treatment.
Transcranial electrical stimulation has widespread clinical and research applications, yet its effect on ongoing neural activity in humans is not well established. Previous reports argue that transcranial alternating current stimulation (tACS) can entrain and enhance neural rhythms related to memory, but the evidence from non-invasive recordings has remained inconclusive. Here, we measure endogenous spindle and theta activity intracranially in humans during low-frequency tACS and find no stable entrainment of spindle power during non-REM sleep, nor of theta power during resting wakefulness. As positive controls, we find robust entrainment of spindle activity to endogenous slow-wave activity in 66% of electrodes as well as entrainment to rhythmic noise-burst acoustic stimulation in 14% of electrodes. We conclude that low-frequency tACS at common stimulation intensities neither acutely modulates spindle activity during sleep nor theta activity during waking rest, likely because of the attenuated electrical fields reaching the cortical surface.
Panmixia and dispersal from the Mediterranean Basin to Macaronesian Islands of a macrolichen species
(2017)
The Mediterranean region, comprising the Mediterranean Basin and the Macaronesian Islands, represents a center of diversification for many organisms. The genetic structure and connectivity of mainland and island microbial populations has been poorly explored, in particular in the case of symbiotic fungi. Here we investigated genetic diversity and spatial structure of the obligate outcrossing lichen-forming fungus Parmelina carporrhizans in the Mediterranean region. Using eight microsatellite and mating-type markers we showed that fungal populations are highly diverse but lack spatial structure. This is likely due to high connectivity and long distance dispersal of fungal spores. Consistent with low levels of linkage disequilibrium and lack of clonality, we detected both mating-type idiomorphs in all populations. Furthermore we showed that the Macaronesian Islands are the result of colonization from the Mediterranean Basin. The unidirectional gene flow, though, seemed not to be sufficient to counterbalance the effects of drift, resulting in comparatively allelic poor peripheral populations. Our study is the first to shed light on the high connectivity and lack of population structure in natural populations of a strictly sexual lichen fungus. Our data further support the view of the Macaronesian Islands as the end of the colonization road for this symbiotic ascomycete.
Colorectal cancer (CRC) has the third highest incidence and the fourth highest mortality rate worldwide and represents a substantial health care burden and affects the life of millions of people. CRC is a genetic disease caused by the stepwise accumulation of genetic alterations. The initiating event in colorectal carcinogenesis is the aberrant activation of the WNT pathway, but other pathways are also commonly deregulated, including the PI3K/AKT pathway. A number of previous studies using genetically engineered mouse models aimed at dissecting the exact role of PI3K/AKT pathway in CRC, but have yielded in rather conflicting results. Despite the inconsistent results, these studies already put forward the idea that PI3K/AKT signaling in combination with other genetic events might substantially contribute to tumor progression.
Since the PI3K/AKT pathway is frequently activated in CRC, it represents an ideal candidate for therapeutic intervention. Although extensive efforts had led to the development of numerous inhibitors targeting the PI3K/AKT pathway, the diversity of genetic alterations can challenge the identification of the most effective therapeutic targets. Therefore, the discovery of shared tumor-promoting mechanisms downstream of these genetic alterations might unravel new biomarkers and druggable targets. The aim of this study was to elucidate the precise role of PI3K/AKT pathway during the course of colorectal carcinogenesis and to decipher novel pro-tumorigenic molecular mechanisms downstream of PI3K/AKT activation that can be used for therapeutic intervention.
To obtain a better insight into the role of the PI3K/AKT pathway during colorectal carcinogenesis, mice expressing an oncogenic variant of AKT1 (AktE17K) specifically in the intestinal epithelial cells (IEC) were used. At the age of 6 months untreated AktE17K mice showed clearly perturbed intestinal homeostasis, but no tumor formation. To induce colonic tumorigenesis, AktE17K mice were subjected to treatment with the colonic carcinogen azoxymethane (AOM). In response to AOM, AktE17K mice developed invasive but nonmetastatic tumors, which showed strong nuclear accumulation of TP53. To investigate the role of PI3K/AKT signaling specifically in CRC progression, AktE17K mice were crossed to TP53- deficient mice (Tp53ΔIEC). Unlike AktE17K mice, untreated Tp53ΔIECAktE17K, developed highly invasive small intestinal tumors by the age of 6 months. To investigate the role of AKT hyperactivation in colonic tumor progression, Tp53ΔIECAktE17K mice were subjected to AOM treatment. AKT hyperactivation significantly enhanced tumor progression and induced metastatic dissemination.
To get a better insight how AKT signaling can promote tumor progression, whole tumor tissues from AOM-treated Tp53ΔIEC and Tp53ΔIECAktE17K mice were subjected to next generation mRNA sequencing and phospho-proteomic analysis by mass spectrometry. Both analyses indicated that AKT hyperactivation expands the inflammatory tumor microenvironment and upregulates pathways associated with invasion and metastasis. Importantly, Gene Set Enrichment Analysis revealed that AOM-induced colon tumors of Tp53ΔIECAktE17K animals, are highly similar in their gene expression profile to the CMS4 subtype of human CRC, which is associated with worse overall- and relapse-free survival7 . Gene expression analysis also suggested elevated NOTCH signaling in the Tp53ΔIECAktE17K tumors. Interestingly, while the expression of Notch3 mRNA was increased in the tumors of Tp53ΔIECAktE17K mice, the expression of the other NOTCH receptors was unaffected by AKT hyperactivation. In vitro experiments using TP53-deficient mouse tumor organoids with hyperactive AKT signaling confirmed the direct, tumor cell-intrinsic link between AKT activation and increased Notch3 expression. Moreover, inhibition of EZH2 mimicked the effect of AKT hyperactivation on Notch3 expression, suggesting that AKT regulates Notch3 via an epigenetic mechanism.
Knock-down of Notch3 in TP53-deficient mouse tumor organoids with hyperactive AKT signaling resulted in differential regulation of several pathways with potential role in invasion and metastasis and in cell death and survival. Subsequent in vivo experiments confirmed the role of NOTCH3 signaling in CRC progression. Treatment of AOM-induced Tp53ΔIECAkt E17K mice with a NOTCH3 antagonistic antibody or the γ-secretase inhibitor DAPT significantly reduced invasion and metastasis. Importantly, NOTCH3 expression was also found to be associated with human CRC progression, suggesting that NOTCH3 represent a valid target for the treatment of CRC. This work, using genetically engineered mouse models and advanced in vitro techniques, has demonstrated a strong tumor promoting role for PI3K/AKT signaling in CRC progression and has identified NOTCH3 signaling as a potential therapeutic target downstream of the PI3K/AKT pathway.
The human brain is one of the most complex biological systems. More than 100 billion neurons build networks that control basic body functions and highly coordinated movements, enable us to express emotions, feelings and thoughts and to store memories over years and even throughout life time. Ultimately, “We are who we are because of what we learn and what we remember” (Kandel 2006). Under pathological conditions, the brain function is challenged. Most if not all neurological diseases have in common that they are either triggered and/or accompanied by inflammatory processes of brain tissue, referred to as neuroinflammation. Such inflammatory processes directly affect an elementary neural mechanism relevant for learning and memory: synaptic plasticity. Indeed, neurons are highly dynamic structures and able to respond to specific stimuli with morphological, functional and molecular adaptations that modify the strength and number of neuronal contact sides (synapses). Hence, the main motivation of this thesis was to identify the neural targets through which inflammation affects brain function and synaptic plasticity in particular. The principles of synaptic plasticity have been studied intensively in the hippocampus, an anatomical structure localized within the temporal lobes that is essential for the consolidation of memories and spatial navigation. Synaptic plasticity is coordinated by complex interactions of thousands of molecules and proteins. Among those proteins, synaptopodin (SP) is localized at a strategic position within excitatory synapses and has been shown to be fundamentally involved in the regulation of synaptic plasticity.
To induce neuroinflammation and to study its effects on SP as well as synaptic plasticity, the classic model of lipopolysaccharide (LPS) was applied. This thesis discloses that inflammatory processes impair the ability of neurons to express hippocampal synaptic plasticity in vivo, which is accompanied by a downregulation of SP-mRNA and protein level in the mouse hippocampus, indicating that SP is one of the cellular targets through which inflammatory signaling pathways affect synaptic plasticity and hence neural function. To learn more about the cellular and molecular mechanisms, an in vitro LPS model was established using entorhino-hippocampal organotypic slice cultures (OTCs).
While confirming the major effect of LPS on SP, this thesis furthermore shows that neuroinflammation crucially involves the cytokine TNFα to transduce its effects on SP, and that microglial cells are the main source of TNFα production under inflammatory conditions. In an attempt to learn more about the mechanisms that are affected under conditions of neuroinflammation effects of retinoic acid (RA), a vitamin A derivate were tested. This is mainly because SP as well as RA have been shown to modulate synaptic plasticity through the accumulation of glutamate receptors at the postsynaptic site: SP via the association with the actincytoskeleton as well as intracellular calcium stores, and RA directly via the modulation of local protein synthesis within dendrites. Indeed, in slice cultures exposed to RA, hippocampal SP cluster size is upregulated, both in vitro and in vivo. Intriguingly, a lack of SP prevents RA-induced synaptic strengthening of hippocampal dentate granule cells in OTCs. This suggests a direct contribution of SP in RA-dependent synaptic plasticity. Interestingly, co-immunoprecipitation of SP-mRNA together with the RA-receptor alpha (RARα) further implies that RA directly controls synaptic plasticity via regulation of SP-protein expression. It is therefore interesting to speculate that RA may increase SP expression or prevent its reduction and thus alterations in synaptic plasticity under conditions of neuroinflammation. Taken together, this thesis identifies SP as an important neuronal target of TNFα-mediated alterations in synaptic plasticity. Moreover, the work on RA indicates that SP affects the ability of neurons to express synaptic plasticity by modulating/mediating local protein synthesis. Since neuroinflammatory processes are an elementary concomitant feature and/or cause of neurological diseases, I am confident that future work on the effects of inflammatory processes on brain function may provide the perspective in devising new therapeutic strategies for the treatment of neuropathologies such as Alzheimer’s disease, multiple sclerosis, epilepsy or stroke, by targeting SP expression and SP-mediated synaptic plasticity.
Pulsed electron-electron double resonance (PELDOR), also called Double Electron-Electron Resonance, (DEER) is a pulsed EPR technique that can provide structural information of biomolecules, such as proteins or nucleic acids, complementary to other structure determination methods by measuring long distances (from 1.5 up to 10 nm) between two paramagnetic labels. Incorporation of the rigid Ç-label pairwise into DNA or RNA molecules enables the determination not only of the distance but also of the mutual orientation between the two Ç-labels by multi-frequency orientation-selective PELDOR data (X-, Q- and G-band frequencies). Thus, information about the orientation of secondary structure elements of nucleic acids can be revealed and used as additional angular information for structure determination. Since Ç does not have motion independent from the helix where it resides, the conformational flexibility of the nucleic acid molecule can be directly determined. This thesis demonstrates the advancement of PELDOR spectroscopy, beyond its original scope of distance measurements, to determine the mutual orientation between two rigid spin labels towards the characterization of the conformational space sampled by highly flexible nucleic acid molecules. Applications of the methodology are shown on two systems: a three-way junction, namely a cocaine aptamer in its bound-state, and a two-way junction, namely a bent DNA.
More in detail, the conformational changes of the cocaine aptamer upon cocaine binding were investigated by analysis of the distance distributions. The cocaine-bound and the unbound states could be differentiated by their conformational flexibility, which decreases in the presence of the ligand. Moreover, the obtained distance distributions revealed a small change in the mean distance between the two spin labels upon cocaine binding. This indicates a ligand-induced conformational change, which presumably originates at the junction where cocaine is known to bind. The investigation of the relative orientation between the two spin-labeled helices of the aptamer revealed further structural insights into the conformational dynamics of the cocaine-bound state. The angular information from the orientation-selective PELDOR data and the a priori knowledge about the secondary structure of the aptamer were helpful in obtaining a molecular model describing its global folding and flexibility. In spite of a large flexible aptamer, the kink angle between the Ç-labeled helices was found to be rather well-defined.
As for the bent DNA molecule, a two-step protocol was proposed to investigate the conformational flexibility. In the first step, a database with all the possible conformers was created, using available restraints from NMR and distance restraints derived from PELDOR. In a second step, a weighted ensemble of these conformers fitting the multi-frequency PELDOR data was built. The uniqueness of the obtained structural ensemble was checked by validation against an independent PELDOR data set recorded at a higher magnetic field strength. In addition, the kink and twist angle pairs were determined and the resulting structural ensemble was compared with the conformational space deduced both from FRET experiments and from the structure determined by the NMR restraints alone.
Overall, this thesis underlines the potential of using PELDOR spectroscopy combined with rigid spin labels in the context of structure determination of nucleic acids in order to determine the relative orientation between two helices, the conformational flexibility and the conformational changes of nucleic acid molecules upon ligand binding.
Algae as primary producers are highly important in aquatic ecosystems and provide a variety of environmental and anthropogenic services. In small lotic ecosystems in agriculturally influenced landscapes, algae are often the main constituent of the base of the food web and they contribute considerably to biodiversity. Within these small lotic ecosystems, algae are influenced by both natural stressors, such as flow regime and dry-out events, and anthropogenic factors. Agricultural practices especially influence algal communities by introducing plant protection products (PPP) and fertilizers into the water. The impacts of these exposures and how they affect planktonic algae in particular are not yet well studied in small lotic ecosystems. However, the protection of algae as primary producers is of high relevance and was thus included in official biomonitoring programs such as the European Water Framework Directive (WFD) or in risk assessment of e.g. PPPs. Hence, this thesis addresses this knowledge gap and links new information on algal communities in small lotic ecosystems with biomonitoring and risk assessment.
Data was gathered from small ditches and streams in central Germany as well as from laboratory algal assays. A technique to rapidly classify and quantify planktonic and benthic algae based on their photopigment concentration (measured via delayed fluorescence - DF) in ecological and ecotoxicological studies was assessed, both in the laboratory and in the field. This research provides insight into planktonic and benthic algal communities in small streams and ditches in order to improve management and protection strategies in the face of increased agricultural chemical input. ...
The challenges posed by climate and land use change are increasingly complex, with ever-increasing and accelerating impacts on the global environmental system. The establishment of an internationally harmonized, integrated, and long-term operated environmental monitoring infrastructure is one of the major challenges of modern environmental research. Increased efforts are currently being made in Europe to establish such a harmonized pan-European observation infrastructure, and the European network of Long-Term Ecological Research sites – LTER-Europe – is of particular importance. By evaluating 477 formally accredited LTER-Europe sites, this study gives an overview of the current distribution of these infrastructures and the present condition of long-term environmental research in Europe. We compiled information on long-term biotic and abiotic observations and measurements and examined the representativeness in terms of continental biogeographical and socio-ecological gradients. The results were used to identify gaps in both measurements and coverage of the aforementioned gradients. Furthermore, an overview of the current state of the LTER-Europe observation strategies is given. The latter forms the basis for investigating the comparability of existing LTER-Europe monitoring concepts both in terms of observational design as well as in terms of the scope of the environmental compartments, variables and properties covered.
Invasive species provide a unique opportunity to evaluate factors controlling biogeographic distributions; we can consider introduction success as an experiment testing suitability of environmental conditions. Predicting potential distributions of spreading species is not easy, and forecasting potential distributions with changing climate is even more difficult. Using the globally invasive coypu (Myocastor coypus [Molina, 1782]), we evaluate and compare the utility of a simplistic ecophysiological based model and a correlative model to predict current and future distribution. The ecophysiological model was based on winter temperature relationships with nutria survival. We developed correlative statistical models using the Software for Assisted Habitat Modeling and biologically relevant climate data with a global extent. We applied the ecophysiological based model to several global circulation model (GCM) predictions for mid-century. We used global coypu introduction data to evaluate these models and to explore a hypothesized physiological limitation, finding general agreement with known coypu distribution locally and globally and support for an upper thermal tolerance threshold. Global circulation model based model results showed variability in coypu predicted distribution among GCMs, but had general agreement of increasing suitable area in the USA. Our methods highlighted the dynamic nature of the edges of the coypu distribution due to climate non-equilibrium, and uncertainty associated with forecasting future distributions. Areas deemed suitable habitat, especially those on the edge of the current known range, could be used for early detection of the spread of coypu populations for management purposes. Combining approaches can be beneficial to predicting potential distributions of invasive species now and in the future and in exploring hypotheses of factors controlling distributions.
Soil frequently occurs as a contaminant on numerous sea, land and air transport pathways. It can carry unwanted invasive species, is widely recognized as a biosecurity risk, and is usually strictly regulated by biosecurity authorities. However, little is known about relative risk levels between pathways, thus authorities have limited capability to identify and target the riskiest soil pathways for management. We conducted a an experiment to test the hypotheses that biosecurity risks from soil organisms will increase both with declining transport duration and with increasing protection from environmental extremes. Soil was collected from two sites, a native forest remnant and an orchard, and stored on, in and under sea containers, or in cupboards, and assayed after 0, 3, 6 and 12 months for bacteria, fungi, nematodes and seeds. Results showed that viability of Pseudomonas spp., bacteria, nematodes and plants declined over 12 months, irrespective of soil source. Also, mortality of most biota was higher when exposed to sunlight, moisture and desiccation than when protected. However, bacterial and fungal numbers were higher in exposed environments, possibly due to ongoing colonization of exposed soil by airborne propagules. The results were consistent with our observations of organisms in soil intercepted from airports and sea ports, and indicated there is potential to rank risks from transported soils based partly on transport duration and environmental exposure. This would help authorities to optimally allocate management resources according to pathway-specific risks.
Introduced species lists provide essential background information for biological invasions research and management. The compilation of these lists is, however, prone to a variety of errors. We highlight the frequency and consequences of such errors using introduced Melaleuca (sensu lato, including Callistemon) species in South Africa as a case study. We examined 111 herbarium specimens from South Africa and noted the categories and sub-categories of errors that occurred in identification. We also used information from herbarium specimens and distribution data collected in the field to determine whether a species was introduced, naturalized and invasive. We found that 72% of the specimens were not named correctly. These were due to human error (70%) (misidentification, and improved identifications) and species identification problems (30%) (synonyms arising from inclusion of Callistemon, and unresolved taxonomy). At least 36 Melaleuca species have been introduced to South Africa, and field observations indicate that ten of these have naturalized, including five that are invasive. While most of the errors likely have negligible impact on management, we highlight one case where incorrect identification lead to an inappropriate management approach and some instances of errors in published lists. Invasive species lists need to be carefully reviewed to minimise errors, and herbarium specimens supported by DNA identification are required where identification using morphological features is particularly challenging.
Biological invasions are occurring frequently and with great impact to agricultural production and other ecosystem services. In response to this, the Australian Weed Risk Assessment (AWRA) was created to assess the potential 'weediness' of plants based on answers to questions related to biogeography, undesirable attributes, and biology or ecology. This basic model has been expanded and adapted for use on other taxa, often without adequate validation. Since invasive insect crop pests are a major economic cost to agricultural production, there is interest in using an expanded model for insects. Here, we review traits related to invasiveness of insects based on a systematic review of the literature. We then compare the identified invasive traits of insects with those identified for plants in the AWRA. Using insects as a case study, we illustrate that although there is some overlap in invasive traits, there are many unique traits related to invasion for both insects and plants. For insects, these traits relate largely to social behaviour. This lack of congruence may also be the case for other taxa. To increase predictive power, a taxon-specific risk assessment tool and deliberate verification are required.
Islands are particularly noteworthy for global conservation because of the high number of species they host, the high levels of species endemism, and the large number and proportion of species at risk of extinction. Much of the conservation threat on islands is from invasive species. Whilst biosecurity is an increasing focus of attention for authorities globally, species are continuing to establish in new locations outside of their native ranges. Among invasive species, ants are a prominent taxon, especially on islands. Over the past decade, following the detection of one of the world’s worst invasive ant species, African big-headed ant Pheidole megacephala, the environmental management authority on world-heritage-listed Lord Howe Island has focused attention on invasive ants. This detection influenced the creation of biosecurity measures to prevent further incursions of exotic species, particularly ants. Despite these efforts, over the following decade numerous ant species were collected on the island for the first time, indicating a serious biosecurity problem. Here, we investigate the chronosequence of ant introductions to Lord Howe Island to quantify the extent and nature of the island’s ant biosecurity problem. A total of 45 species have been collected on the island and of these, 12 are considered to be endemic, and a further seven are possibly native. Nineteen of the 26 introduced species (42% of the total fauna and 73% of the introduced fauna) were only found for the first time in the last 15 years. All but two of the species that are not native to Lord Howe Island are native to the Australian mainland, indicating that the biosecurity threat comes from the transport of goods from the Australian mainland. We suggest that the pattern of accelerating ant species accumulation on Lord Howe Island is probably not an isolated phenomenon, and that it is probably occurring on most islands globally that are habitable by ants and visited by people.
Factors that cause differential establishment among naturalized, invasive, and native species are inadequately documented, much less often quantified among different communities. We evaluated the effects of seed addition and disturbance (i.e., understory canopy removal) on the establishment and seedling biomass among two naturalized, two invasive, and two native species (1 forb, 1 grass in each group) within steppe and low elevation forest communities in eastern Washington, USA. Establishment within each plant immigrant class was enhanced by seed addition: naturalized species showed the greatest difference in establishment between seed addition and no seed addition plots, native and invasive species establishment also increased following seed addition but not to the same magnitude as naturalized species. Within seed addition plots, understory canopy disturbance resulted in significant increases in plant establishment (regardless of plant immigration class) relative to undisturbed plots and the magnitude of this effect was comparable between steppe and adjacent forest. However, regardless of disturbance treatment fewer invasive plants established in the forest than in the steppe, whereas native and naturalized plant establishment did not differ between the habitats. Individual biomass of naturalized species were consistently greater in disturbed (canopy removed) versus undisturbed control plots and naturalized species were also larger in the steppe than in the forest at the time of harvest. Similar trends in plant size were observed for the native and invasive species, but the differences in biomass for these two immigration classes between disturbance treatments and between habitats were not significant. We found that strong limitations of non-native species is correlated with intact canopy cover within the forest understory, likely driven by the direct or indirect consequences of low light transmittance through the arboreal and understory canopy. Considered collectively, our results demonstrate how seed limitation and intact plant ground cover can limit the abundance and performance of naturalized species in Pacific Northwest steppe and low elevation forest, suggesting that local disturbance in both habitats creates microsites for these species to establish and survive. Future studies evaluating interactions between multiple barriers to establishment using more representatives from each immigration class will further reveal how biotic interactions ultimately influence the demography and distribution of non-native plants within these communities.
In the face of increasing invasions and limited resources, appropriate management of invasive species requires prioritisation of species for management action. This process often relies on knowledge of species specific impacts. However, as studies explicitly measuring impact of marine alien species are rare, prioritisation of management actions is often based on studies from outside the geographic area of interest. Further, few impact studies account for context dependency (e.g. seasonal variability or distinct environmental regimes), raising the question of how transferrable knowledge about the impact of a species is between invaded ranges. This study addressed this question by using the widespread invasive solitary ascidian Ciona robusta as a case study for assessing impacts across two invaded regions: South Africa and California, USA. We replicated a previously conducted experiment from California that showed that C. robusta depresses local species richness in San Francisco Bay. Our South African experiment showed no effect of C. robusta on species richness, the Shannon-Weiner diversity index or community composition, despite experiments being carried out over two years and at two depths. While these results may reflect strong density dependency in the impact of C. robusta, they serve to highlight context dependency in invasive species impacts. This suggests that until studies of impact in marine systems become common place, context dependency should be explicitly addressed as a source of uncertainty during the prioritisation of species for management action.
Carbon mass of the non-indigenous predatory fishhook water flea Cercopagis pengoi (Ostroumov, 1891) from the eastern Gulf of Finland, the Baltic Sea, was for the first time measured using the high temperature combustion method. Prior to the analysis, individual dry weight of Cercopagis was determined; altogether ca. 500 organisms were examined. Mean individual dry weight of C. pengoi for July-September was estimated as 34.0 μg; carbon mass averaged 15.8 μg; carbon content, calculated as percent of dry weight, averaged 43.4%. Those values varied over months, mainly because of different population structure of C. pengoi and variation in their diet due to seasonal dynamics of the food objects. However, relations between carbon mass and dry weight for different months did not differ statistically (p<0.001). Therefore, the general polynomial regressions (k=2), describing carbon mass-to-dry weight and carbon content-to-dry weight relationships, were calculated for the entire dataset of individual measurements of C. pengoi body metrics. These data will contribute to adequate evaluation of food web structure and ecosystem alterations in various water bodies invaded by C. pengoi which has got a strong potential to pelagic food web transformations that may impact the overall energy balance and decrease the size of fish stocks.
Prioritization of introduction pathways is seen as an important component of the management of biological invasions. We address whether established alien plants, mammals, freshwater fish and terrestrial invertebrates with known ecological impacts are associated with particular introduction pathways (release, escape, contaminant, stowaway, corridor and unaided). We used the information from the European alien species database DAISIE (www.europe-aliens.org) supplemented by the EASIN catalogue (European Alien Species Information Network), and expert knowledge. Plants introduced by the pathways release, corridor and unaided were disproportionately more likely to have ecological impacts than those introduced as contaminants. In contrast, impacts were not associated with particular introduction pathways for invertebrates, mammals or fish. Thus, while for plants management strategies should be targeted towards the appropriate pathways, for animals, management should focus on reducing the total number of taxa introduced, targeting those pathways responsible for high numbers of introductions. However, regardless of taxonomic group, having multiple introduction pathways increases the likelihood of the species having an ecological impact. This may simply reflect that species introduced by multiple pathways have high propagule pressure and so have a high probability of establishment. Clearly, patterns of invasion are determined by many interacting factors and management strategies should reflect this complexity.
Humans have an extremely long history of transporting and introducing mammal species outside their native geographic ranges. The characteristics of the species introduced (taxonomy, life-history, ecology, environment) can all influence which traits are available (and selected) for establishment, and subsequent invasive spread. Understanding the non-randomness in species introductions is therefore key to understanding invasions by alien species. Here, we test for selectivity in the identities and traits of mammal species introduced worldwide. We compiled and analysed a comprehensive database of introduced mammal species, including information on a broad range of life history, ecological, distributional and environmental variables that we predicted to differ between introduced and non-introduced mammal species. Certain mammal taxa are much more likely to have been introduced than expected, such as Artiodactyls in the families Bovidae and Cervidae. Rodents and bats were much less likely to have been introduced than expected. Introduced mammal species have significantly larger body masses, longer lifespans and larger litter sizes than a random sample of all mammal species. They also have much larger native geographic ranges than expected, originate from significantly further north, from cooler areas, and from areas with higher human population densities, than mammal species with no recorded introductions. The traits and distributions of species help determine which have been introduced, and reflect how the evolutionary history of mammals has resulted in certain species with certain traits being located in the way of human histories of movement and demands for goods and services. The large amount of unexplained variation is likely to relate to the intrinsically stochastic nature of this human-driven process.
Impact assessment with different scoring tools: How well do alien amphibian assessments match?
(2017)
Classification of alien species' impacts can aid policy making through evidence based listing and management recommendations. We highlight differences and a number of potential difficulties with two scoring tools, the Environmental Impact Classification of Alien Taxa (EICAT) and the Generic Impact Scoring System (GISS) using amphibians as a case study. Generally, GISS and EICAT assessments lead to very similar impact levels, but scores from the schemes are not equivalent. Small differences are attributable to discrepancies in the verbal descriptions for scores. Differences were found in several impact categories. While the issue of disease appears to be related to uncertainties in both schemes, hybridisation might be inflated in EICAT. We conclude that GISS scores cannot directly be translated into EICAT classifications, but they give very similar outcomes and the same literature base can be used for both schemes.
Range expansion drives the evolution of alternate reproductive strategies in invasive fire ants
(2017)
Many species are expanding their ranges in response to climate changes or species introductions. Expansion-related selection likely drives the evolution of dispersal and reproductive traits, especially in invasive species introduced into novel habitats. We used an agent-based model to investigate these relationships in the red imported fire ant, Solenopsis invicta, by tracking simulated populations over 25 years. Most colonies of this invasive species produce two types of queens practicing alternate reproductive strategies. Claustral queens found new colonies in vacant habitats, while parasitic queens take over existing colonies whose queens have died. We investigated how relative investment in the two queen types affects population demography, habitat occupancy, and range expansion. We found that parasitic queens extend the ecological lifespan of colonies, thereby increasing a population’s overall habitat occupancy as well as average colony size (number of workers) and territory size. At the same time, investment in parasitic queens slowed the rate of range expansion by diverting investment from claustral queens. Divergent selection regimes caused edge and interior populations to evolve different levels of reproductive investment, such that interior populations invested heavily in parasitic queens whereas those at the edge invested almost entirely in claustral queens. Our results highlight factors shaping ant life histories, including the evolution of social parasitism, and have implications for the response of species to range shifts.
Plant traits are critical for understanding invasion success of introduced species, yet attempts to identify universal traits that explain invasion success and impact have been unsuccessful because environmenttrait- fitness relationships are complex, potentially context dependent, and variation in traits is often unaccounted for. As introduced species encounter novel environments, their traits and trait variability can determine their ability to grow and reproduce, yet invasion biologists do not often have an understanding of how novel environments might shape traits. To uncover which combination of traits are most effective for predicting invasion success, we studied three different urban habitat types along the Nile Delta in Egypt invaded by the Pink Morning Glory, Ipomoea carnea Jacq. (Family: Convolvulaceae). Over two years, we measured ten plant traits at monthly intervals along an invasion gradient in each habitat. No single trait sufficiently explained survival probability and that traits linked to invasion success were better predicted by the characteristics of the invaded habitat. While the measured traits did influence survival of I. carnea, the importance of specific traits was contingent on the local environment, meaning that local trait-environment interactions need to be understood in order to predict invasion.
The adoption of a consistent alien species pathways categorization, hierarchy and terminology is crucial for increasing the interoperability of different online databases. In the present paper the European Alien Species Information Network (EASIN) classification system of pathways is compared and discussed with the classification scheme recently published by the Convention on Biological Diversity (CBD). Although the main pathway categories of the two classifications overall match, there are substantial differences in their subcategorization, with EASIN including 20 pathway subcategories while CBD considers 44 subcategories. In most cases, each EASIN subcategory pathway can correspond to two or more CBD subcategories. About 5,500 species listed in EASIN do not match directly with the CBD pathway subcategories, most of which are terrestrial invertebrates. Aiming at achieving synchronization between the two classification systems and at facilitating the access to information to researchers and policy makers, EASIN is trying to align with the CBD pathway classification scheme. This alignment process requires the involvement of a large number of experts, especially from the terrestrial realm, and the adoption of commonly accepted definitions of the CBD pathways.
Thirty-seven alien plant species, pre-identified by horizon scanning exercises were prioritised for pest risk analysis (PRA) using a modified version of the EPPO Prioritisation Process designed to be compliant with the EU Regulation 1143/2014. In Stage 1, species were categorised into one of four lists – a Residual List, EU List of Minor Concern, EU Observation List and the EU List of Invasive Alien Plants. Only those species included in the latter proceeded to the risk management stage where their priority for PRA was assessed. Due to medium or high spread potential coupled with high impacts twenty-two species were included in the EU List of Invasive Alien Plants and proceeded to Stage 2. Four species (Ambrosia trifida, Egeria densa, Fallopia baldschuanica and Oxalis pes-caprae) were assigned to the EU Observation List due to moderate or low impacts. Albizia lebbeck, Clematis terniflora, Euonymus japonicus, Lonicera morrowii, Prunus campanulata and Rubus rosifolius were assigned to the residual list due to a current lack of information on impacts. Similarly, Cornus sericea and Hydrilla verticillata were assigned to the Residual List due to unclear taxonomy and uncertainty in native status, respectively. Chromolaena odorata, Cryptostegia grandiflora and Sphagneticola trilobata were assigned to the Residual List as it is unlikely they will establish in the Union under current climatic conditions. In the risk management stage, Euonymus fortunei, Ligustrum sinense and Lonicera maackii were considered a low priority for PRA as they do not exhibit invasive tendencies despite being widely cultivated in the EU over several decades. Nineteen species were identified as having a high priority for a PRA (Acacia dealbata, Ambrosia confertiflora, Andropogon virginicus, Cardiospermum grandiflorum, Celastrus orbiculatus, Cinnamomum camphora, Cortaderia jubata, Ehrharta calycina, Gymnocoronis spilanthoides, Hakea sericea, Humulus scandens, Hygrophila polysperma, Lespedeza cuneata, Lygodium japonicum, Pennisetum setaceum, Prosopis juliflora, Sapium sebiferum, Pistia stratiotes and Salvinia molesta).
The value of plant ecological datasets with hundreds or thousands of species is principally determined by the taxonomic accuracy of their plant names. However, combining existing lists of species to assemble a harmonized dataset that is clean of taxonomic errors can be a difficult task for non-taxonomists. Here, we describe the range of taxonomic difficulties likely to be encountered during dataset assembly and present an easy-to-use taxonomic cleaning protocol aimed at assisting researchers not familiar with the finer details of taxonomic cleaning. The protocol produces a final dataset (FD) linked to a companion dataset (CD), providing clear details of the path from existing lists to the FD taken by each cleaned taxon. Taxa are checked off against ten categories in the CD that succinctly summarize all taxonomic modifications required. Two older, publicly-available lists of naturalized Asteraceae in Australia were merged into a harmonized dataset as a case study to quantify the impacts of ignoring the critical process of taxonomic cleaning in invasion ecology. Our FD of naturalized Asteraceae contained 257 species and infra-species. Without implementation of the full cleaning protocol, the dataset would have contained 328 taxa, a 28% overestimate of taxon richness by 71 taxa. Our naturalized Asteraceae CD described the exclusion of 88 names due to nomenclatural issues (e.g. synonymy), the inclusion of 26 updated currently accepted names and four taxa newly naturalized since the production of the source datasets, and the exclusion of 13 taxa that were either found not to be in Australia or were in fact doubtfully naturalized. This study also supports the notion that automated processes alone will not be enough to ensure taxonomically clean datasets, and that manual scrutiny of data is essential. In the long term, this will best be supported by increased investment in taxonomy and botany in university curricula.